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Depletion of nuclear LINE1 RNA in mouse ESCs and embryos

LINE1 is the most active and abundant family of retrotransposons; it is implicated in a number of pathologies, as well as in early embryo development. We present a protocol to specifically knockdown LINE1 in mouse embryonic stem cells and embryos, including details for the nucleofection and zygote m...

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Autores principales: Percharde, Michelle, Lin, Chih-Jen, Ramalho-Santos, Miguel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8361322/
https://www.ncbi.nlm.nih.gov/pubmed/34409306
http://dx.doi.org/10.1016/j.xpro.2021.100726
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author Percharde, Michelle
Lin, Chih-Jen
Ramalho-Santos, Miguel
author_facet Percharde, Michelle
Lin, Chih-Jen
Ramalho-Santos, Miguel
author_sort Percharde, Michelle
collection PubMed
description LINE1 is the most active and abundant family of retrotransposons; it is implicated in a number of pathologies, as well as in early embryo development. We present a protocol to specifically knockdown LINE1 in mouse embryonic stem cells and embryos, including details for the nucleofection and zygote microinjection of LINE antisense oligos, followed by RNA FISH validation. This protocol can be used in development, as well as other cell types where LINE1 is believed to be expressed. For complete information on the use and execution of this protocol, please refer to Percharde et al. (2018).
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spelling pubmed-83613222021-08-17 Depletion of nuclear LINE1 RNA in mouse ESCs and embryos Percharde, Michelle Lin, Chih-Jen Ramalho-Santos, Miguel STAR Protoc Protocol LINE1 is the most active and abundant family of retrotransposons; it is implicated in a number of pathologies, as well as in early embryo development. We present a protocol to specifically knockdown LINE1 in mouse embryonic stem cells and embryos, including details for the nucleofection and zygote microinjection of LINE antisense oligos, followed by RNA FISH validation. This protocol can be used in development, as well as other cell types where LINE1 is believed to be expressed. For complete information on the use and execution of this protocol, please refer to Percharde et al. (2018). Elsevier 2021-08-09 /pmc/articles/PMC8361322/ /pubmed/34409306 http://dx.doi.org/10.1016/j.xpro.2021.100726 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Percharde, Michelle
Lin, Chih-Jen
Ramalho-Santos, Miguel
Depletion of nuclear LINE1 RNA in mouse ESCs and embryos
title Depletion of nuclear LINE1 RNA in mouse ESCs and embryos
title_full Depletion of nuclear LINE1 RNA in mouse ESCs and embryos
title_fullStr Depletion of nuclear LINE1 RNA in mouse ESCs and embryos
title_full_unstemmed Depletion of nuclear LINE1 RNA in mouse ESCs and embryos
title_short Depletion of nuclear LINE1 RNA in mouse ESCs and embryos
title_sort depletion of nuclear line1 rna in mouse escs and embryos
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8361322/
https://www.ncbi.nlm.nih.gov/pubmed/34409306
http://dx.doi.org/10.1016/j.xpro.2021.100726
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